Method of treating a bacterial infection using colostrum

ABSTRACT

The method of treating a bacterial infection using colostrum includes administering an effective amount of colostrum to a subject in need thereof. The infection can be caused by G+ or G− bacteria. The colostrum administered may be selected from the group consisting of bovine colostrum, camel colostrum, and a mixture of bovine colostrum and camel colostrum. The bacterial infection may be selected from the group consisting of Staphylococcus aureus subs. aureus Rosenbach, Escherichia coli, Pseudomonas aeruginosa, and Methicillin-resistant Staphylococcus aureus. A colostrum composition can include a mixture of bone and camel colostrum and a pharmaceutically acceptable carrier.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a division of Ser. No. 16/809,882, filed Mar. 5,2020, pending, the priority of which is claimed.

BACKGROUND 1. Field

The disclosure of the present patent application relates to a method oftreating a bacterial infection by inducing an immune response.

2. Description of the Related Art

The CDC identifies antibiotic resistance as one of the most significantpublic health challenges of our time. In the United States alone, atleast 2 million people are diagnosed with antibiotic-resistantinfections annually, and at least 23,000 of these are fatal.

Strategies for developing new treatments have mostly focused on avoidingthe rise of antibiotic resistance and on developing new antibiotics.However, avoiding the rise of antibiotic resistance is a failingrearguard action, given the prevalence of antibiotic-resistant bacterialstrains. Further, the rate of new antibiotic development has slowedsignificantly. Most new classes of antibiotics were developed in the1940s and the 1950s, with only two new classes developed since the year2000. Antibiotics also have many undesirable side effects, ranging fromnausea and diarrhea to liver and kidney damage.

Thus, a method of treating an infection solving the aforementionedproblems is desired.

SUMMARY

A method of treating a bacterial infection using mixed colostrum caninclude administering a therapeutically effective amount of colostrum toa patient in need thereof. Administering colostrum to the patient caninduce an immune response in the patient. In an embodiment, thecolostrum administered may be selected from the group consisting ofbovine colostrum, camel colostrum, and a mixture of bovine colostrum andcamel colostrum. The infection can be bacterial. In an embodiment, thebacterial infection may be selected from the group consisting of G+ andG− bacteria: Staphylococcus aureus subs. aureus Rosenbach, Escherichiacoli, Pseudomonas aeruginosa, and Methicillin-resistant Staphylococcusaureus.

An embodiment of the present subject matter is directed to apharmaceutical composition including the mixture of bovine and camelcolostrum and a pharmaceutically acceptable carrier.

An embodiment of the present subject matter is directed to a method ofmaking a pharmaceutical composition including mixing the colostrum understerile conditions with a pharmaceutically acceptable carrier andpreservatives, buffers, or propellants to create the pharmaceuticalcomposition; and providing the pharmaceutical composition in a formsuitable for daily, weekly, or monthly administration. In an embodiment,the pharmaceutical composition may be natural and/or organic, comprisingexclusively natural and/or organic ingredients

An embodiment of the present subject matter is directed to a method oftreating a bacterial infection, including administering to a subject inneed thereof a therapeutically effective amount of a pharmaceuticalcomposition according to the present subject matter. The pharmaceuticalcomposition can include a mixture of bovine and camel colostrum. In anembodiment, the pharmaceutical composition may be an organicpharmaceutical composition.

These and other features of the present disclosure will become readilyapparent upon further review of the following specification anddrawings.

BRIEF DESCRIPTION OF THE DRAWINGS

The patent or application file contains at least one drawing executed incolor. Copies of this patent or patent application publication withcolor drawing(s) will be provided by the Office upon request and paymentof the necessary fee.

FIG. 1 depicts a bar graph displaying IFN-γ levels in control subjects,subjects exposed to colostrum alone, subjects exposed to E. coli alone,and subjects exposed to both E. coli and colostrum, where the colostrumwas either bovine colostrum, camel colostrum, or a mixture of bovine andcamel colostrum.

FIG. 2A depicts a bar graph displaying IFN-γ levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to E.coli alone, and subjects exposed to both E. coli and colostrum, wherethe colostrum was camel colostrum.

FIG. 2B depicts a bar graph displaying IFN-γ levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to E.coli alone, and subjects exposed to both E. coli and colostrum, wherethe colostrum was bovine colostrum.

FIG. 2C depicts a bar graph displaying IFN-γ levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to E.coli alone, and subjects exposed to both E. coli and colostrum, wherethe colostrum was a mixture of camel colostrum and bovine colostrum.

FIG. 3 depicts a bar graph displaying TNF-α levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to E.coli alone, and subjects exposed to both E. coli and colostrum, wherethe colostrum was bovine, camel, or a mixture of bovine colostrum andcamel colostrum.

FIG. 4A depicts a bar graph displaying TNF-α levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to E.coli alone, and subjects exposed to both E. coli and colostrum, wherethe colostrum was camel colostrum.

FIG. 4B depicts a bar graph displaying TNF-α levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to E.coli alone, and subjects exposed to both E. coli and colostrum, wherethe colostrum was bovine colostrum.

FIG. 4C depicts a bar graph displaying TNF-α levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to E.coli alone, and subjects exposed to both E. coli and colostrum, wherethe colostrum was a mixture of camel colostrum and bovine colostrum.

FIG. 5 depicts a bar graph displaying IL-10 levels in control subjects,subjects exposed to colostrum alone, subjects exposed to E. coli alone,and subjects exposed to both E. coli and colostrum, where the colostrumwas either bovine colostrum, camel colostrum, or a mixture of bovinecolostrum and camel colostrum.

FIG. 6A depicts a bar graph displaying IL-10 levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to E.coli alone, and subjects exposed to both E. coli and colostrum, wherethe colostrum was camel colostrum.

FIG. 6B depicts a bar graph displaying IL-10 levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to E.coli alone, and subjects exposed to both E. coli and colostrum, wherethe colostrum was bovine colostrum.

FIG. 6C depicts a bar graph displaying IL-10 levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to E.coli alone, and subjects exposed to both E. coli and colostrum, wherethe colostrum was a mixture of camel colostrum and bovine colostrum.

FIG. 7 depicts a bar graph displaying IFN-γ levels in control subjects,subjects exposed to colostrum alone, subjects exposed to P. aeruginosaalone, and subjects exposed to both P. aeruginosa and colostrum, wherethe colostrum was either bovine colostrum, camel colostrum, or a mixtureof bovine colostrum and camel colostrum.

FIG. 8A depicts a bar graph displaying IFN-γ levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to P.aeruginosa alone, and subjects exposed to both P. aeruginosa andcolostrum, where the colostrum was camel colostrum.

FIG. 8B depicts a bar graph displaying IFN-γ levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to P.aeruginosa alone, and subjects exposed to both P. aeruginosa andcolostrum, where the colostrum was bovine colostrum.

FIG. 8C depicts a bar graph displaying IFN-γ levels over time in controlsubjects, subjects exposed to colostrum alone, subjects exposed to P.aeruginosa alone, and subjects exposed to both P. aeruginosa andcolostrum, where the colostrum was a mixture of camel colostrum andbovine colostrum.

FIG. 9 depicts a bar graph displaying TNF-α levels in control subjects,subjects exposed to colostrum alone, subjects exposed to P. aeruginosaalone, and subjects exposed to both P. aeruginosa and colostrum, wherethe colostrum was either bovine colostrum, camel colostrum, or a mixtureof bovine colostrum and camel colostrum.

FIG. 10A depicts a bar graph displaying TNF-α levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto P. aeruginosa alone, and subjects exposed to both P. aeruginosa andcolostrum, where the colostrum was camel colostrum.

FIG. 10B depicts a bar graph displaying TNF-α levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto P. aeruginosa alone, and subjects exposed to both P. aeruginosa andcolostrum, where the colostrum was bovine colostrum.

FIG. 10C depicts a bar graph displaying TNF-α levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto P. aeruginosa alone, and subjects exposed to both P. aeruginosa andcolostrum, where the colostrum was a mixture of camel colostrum andbovine colostrum.

FIG. 11 depicts a bar graph displaying IL-10 levels in control subjects,subjects exposed to colostrum alone, subjects exposed to P. aeruginosaalone, and subjects exposed to both P. aeruginosa and colostrum, wherethe colostrum was either bovine colostrum, camel colostrum, or a mixtureof bovine colostrum and camel colostrum.

FIG. 12A depicts a bar graph displaying IL-10 levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto P. aeruginosa alone, and subjects exposed to both P. aeruginosa andcolostrum, where the colostrum was camel colostrum.

FIG. 12B depicts a bar graph displaying IL-10 levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto P. aeruginosa alone, and subjects exposed to both P. aeruginosa andcolostrum, where the colostrum was bovine colostrum.

FIG. 12C depicts a bar graph displaying IL-10 levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto P. aeruginosa alone, and subjects exposed to both P. aeruginosa andcolostrum, where the colostrum was a mixture of camel colostrum andbovine colostrum.

FIG. 13 depicts a bar graph displaying IFN-γ levels in control subjects,subjects exposed to colostrum alone, subjects exposed to S. aureusalone, and subjects exposed to both S. aureus and colostrum, where thecolostrum was either bovine colostrum, camel colostrum, or a mixture ofbovine colostrum and camel colostrum.

FIG. 14A depicts a bar graph displaying IFN-γ levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto S. aureus alone, and subjects exposed to both S. aureus andcolostrum, where the colostrum was camel colostrum.

FIG. 14B depicts a bar graph displaying IFN-γ levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto S. aureus alone, and subjects exposed to both S. aureus andcolostrum, where the colostrum was bovine colostrum.

FIG. 14C depicts a bar graph displaying IFN-γ levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto S. aureus alone, and subjects exposed to both S. aureus andcolostrum, where the colostrum was a mixture of camel colostrum andbovine colostrum.

FIG. 15 depicts a bar graph displaying TNF-α levels in control subjects,subjects exposed to colostrum alone, subjects exposed to S. aureusalone, and subjects exposed to both S. aureus and colostrum, where thecolostrum was either bovine colostrum, camel colostrum, or a mixture ofbovine colostrum and camel colostrum.

FIG. 16A depicts a bar graph displaying TNF-α levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto S. aureus alone, and subjects exposed to both S. aureus andcolostrum, where the colostrum was camel colostrum.

FIG. 16B depicts a bar graph displaying TNF-α levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto S. aureus alone, and subjects exposed to both S. aureus andcolostrum, where the colostrum was bovine colostrum.

FIG. 16C depicts a bar graph displaying TNF-α levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto S. aureus alone, and subjects exposed to both S. aureus andcolostrum, where the colostrum was a mixture of camel colostrum andbovine colostrum.

FIG. 17 depicts a bar graph displaying IL-10 levels in control subjects,subjects exposed to colostrum alone, subjects exposed to S. aureusalone, and subjects exposed to both S. aureus and colostrum, where thecolostrum was either bovine colostrum, camel colostrum, or a mixture ofbovine colostrum and camel colostrum.

FIG. 18A depicts a bar graph displaying IL-10 levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto S. aureus alone, and subjects exposed to both S. aureus andcolostrum, where the colostrum was camel colostrum.

FIG. 18B depicts a bar graph displaying IL-10 levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto S. aureus alone, and subjects exposed to both S. aureus andcolostrum, where the colostrum was bovine colostrum.

FIG. 18C depicts a bar graph displaying IL-10 levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto S. aureus alone, and subjects exposed to both S. aureus andcolostrum, where the colostrum was a mixture of camel colostrum andbovine colostrum.

FIG. 19 depicts a bar graph displaying IFN-γ levels in control subjects,subjects exposed to colostrum alone, subjects exposed to MRSA, andsubjects exposed to both MRSA and colostrum, where the colostrum waseither bovine colostrum, camel colostrum, or a mixture of bovinecolostrum and camel colostrum.

FIG. 20A depicts a bar graph displaying IFN-γ levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto MRSA alone, and subjects exposed to both MRSA and colostrum, wherethe colostrum was camel colostrum.

FIG. 20B depicts a bar graph displaying IFN-γ levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto MRSA alone, and subjects exposed to both MRSA and colostrum, wherethe colostrum was bovine colostrum.

FIG. 20C depicts a bar graph displaying IFN-γ levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto MRSA alone, and subjects exposed to both MRSA and colostrum, wherethe colostrum was a mixture of camel colostrum and bovine colostrum.

FIG. 21 depicts a bar graph displaying TNF-α levels in control subjects,subjects exposed to colostrum alone, subjects exposed to MRSA, andsubjects exposed to both MRSA and colostrum, where the colostrum waseither bovine colostrum, camel colostrum, or a mixture of bovinecolostrum and camel colostrum.

FIG. 22A depicts a bar graph displaying TNF-α levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto MRSA alone, and subjects exposed to both MRSA and colostrum, wherethe colostrum was camel colostrum.

FIG. 22B depicts a bar graph displaying TNF-α levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto MRSA alone, and subjects exposed to both MRSA and colostrum, wherethe colostrum was bovine colostrum.

FIG. 22C depicts a bar graph displaying TNF-α levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto MRSA alone, and subjects exposed to both MRSA and colostrum, wherethe colostrum was a mixture of camel colostrum and bovine colostrum.

FIG. 23 depicts a bar graph displaying IL-10 levels in control subjects,subjects exposed to colostrum alone, subjects exposed to MRSA, andsubjects exposed to both MRSA and colostrum, where the colostrum waseither bovine colostrum, camel colostrum, or a mixture of bovinecolostrum and camel colostrum.

FIG. 24A depicts a bar graph displaying IL-10 levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto MRSA alone, and subjects exposed to both MRSA and colostrum, wherethe colostrum was camel colostrum.

FIG. 24B depicts a bar graph displaying IL-10 levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto MRSA alone, and subjects exposed to both MRSA and colostrum, wherethe colostrum was bovine colostrum.

FIG. 24C depicts a bar graph displaying IL-10 levels over time incontrol subjects, subjects exposed to colostrum alone, subjects exposedto MRSA alone, and subjects exposed to both MRSA and colostrum, wherethe colostrum was a mixture of camel colostrum and bovine colostrum.

Similar reference characters denote corresponding features consistentlythroughout the attached drawings.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

A method of treating a bacterial infection may include administeringcolostrum to a subject in need thereof. In an embodiment, the colostrumadministered may be selected from the group consisting of bovinecolostrum, camel colostrum, and a mixture of bovine colostrum and camelcolostrum. In an embodiment, the infection is a bacterial infection oran infection caused by bacteria. The bacterial infection may be selectedfrom the group consisting of G+ bacteria and G− bacteria. The bacterialinfection may be selected from the group consisting of Staphylococcusaureus subs. aureus Rosenbach (S. aureus), Escherichia coli (E. coli),Pseudomonas aeruginosa (P. aeruginosa), and Methicillin-resistantStaphylococcus aureus (MRSA).

As used herein, a “subject” includes mammals, e.g., humans, dogs, cats,sheep, cows, rats, mice, and the like.

As used herein, the term “about,” when used to modify a numerical value,means within ten percent of that numerical value.

As used herein, “colostrum” is the first form of milk produced by themammary gland of a mammal just before and/or shortly after giving birth.Colostrum contains maternal antibodies intended to protect the newbornagainst disease, and frequently contains significantly more protein thannon-colostrum milk.

In an embodiment, the colostrum may be collected after parturition andstored at −20° C. for at least a week. In an embodiment, the colostrummay then be thawed at −4° C. for at least 8 hours, and then lyophilized.In an embodiment, the lyophilized colostrum may then be suspended insterilized saline solution at a concentration of 28 g lyophilizedcolostrum per liter of sterilized saline solution, forming a colostrumcomposition for use as discussed herein. In an embodiment, a colostrumcomposition may include bovine colostrum, camel colostrum, or a mixtureof bovine colostrum and camel colostrum. An embodiment of the presentsubject matter may include the colostrum composition including at leasta mixture of one of the bovine colostrum and the camel colostrum and apharmaceutically acceptable carrier.

In an embodiment, the administration of colostrum or a colostrumcomposition of the present subject matter may include dailyadministration. In an embodiment, the daily administration may beadministered in the early morning hours. In an embodiment,administration of colostrum or a colostrum composition of the presentsubject matter may include injecting one ml of a solution of 28 glyophilized colostrum per liter of sterilized saline solution every dayat eight in the morning for at least a month.

In an embodiment, administration of the colostrum or colostrumcomposition as described herein may prevent the lethal effect of alethal dose of an infectious bacterium. In an embodiment, the infectiousbacterium may be selected from the group consisting of E. coli, P.aeruginosa, S. aureus, and MRSA.

In an embodiment, administration of the colostrum or colostrumcomposition of the present subject matter may stimulate the immunesystem. In an embodiment, the immune system stimulation may include anincrease in the level of one or more cytokines in the blood. In anembodiment, the cytokines may be selected from the group consisting ofInterferon-7 (IFN-γ), Tumor Necrosis Factor α (TNF-α), andInterleukin-10 (IL-10).

An embodiment of the present subject matter is directed to apharmaceutical composition comprising the mixture of bovine and camelcolostrum and a pharmaceutically acceptable carrier.

An embodiment of the present subject matter is directed to a method ofmaking a pharmaceutical composition, including mixing an effectiveamount of the bovine and camel mixture colostrum with a pharmaceuticallyacceptable carrier. For example, the method of making a pharmaceuticalcomposition can include mixing the colostrum under sterile conditionswith a pharmaceutically acceptable carrier, preservatives, buffers,and/or propellants to create the pharmaceutical composition. In anembodiment, the pharmaceutical composition may be natural and/or organicand the pharmaceutically acceptable carrier may also be naturaland/organic.

To prepare the pharmaceutical composition, at least one of the bovinecolostrum and camel colostrum, as the active ingredient, is intimatelyadmixed with a pharmaceutically acceptable carrier according toconventional pharmaceutical compounding techniques. Carriers are inertpharmaceutical excipients, including, but not limited to, binders,suspending agents, lubricants, flavorings, sweeteners, preservatives,dyes, and coatings. In preparing compositions in oral dosage form, anyof the pharmaceutical carriers known in the art may be employed. Forexample, for liquid oral preparations, suitable carriers and additivesinclude water, glycols, oils, alcohols, flavoring agents, preservatives,coloring agents, and the like. Further, for solid oral preparations,suitable carriers and additives include starches, sugars, diluents,granulating agents, lubricants, binders, disintegrating agents, and thelike.

The present compositions can be in unit dosage forms such as tablets,pills, capsules, powders, granules, ointments, sterile parenteralsolutions or suspensions, metered aerosol or liquid sprays, drops,ampules, auto-injector devices or suppositories, for oral parenteral,intranasal, sublingual or rectal administration, or for administrationby inhalation or insufflation. The active compound can be mixed understerile conditions with a pharmaceutically acceptable carrier and, ifrequired, any needed preservatives, buffers, or propellants. Thecomposition can be presented in a form suitable for daily, weekly, ormonthly administration. The pharmaceutical compositions herein willcontain, per dosage unit, e.g., tablet, capsule, powder, injection,teaspoonful, suppository and the like, an amount of the activeingredient necessary to deliver an effective dose. A therapeuticallyeffective amount of the colostrum compositions or an amount effective totreat a disease, such as a bacterial infection, may be determinedinitially from the Examples described herein and adjusted for specifictargeted diseases using routine methods.

The colostrum or colostrum compositions can be administered to a subjectin need thereof. For example, the colostrum or colostrum compositionscan be used to treat a subject suffering from a bacterial infection. Thebacterial infection can be caused by E. coli, P. aeruginosa, S. aureus,MRSA, or the like.

An embodiment of the present subject matter is directed to a method oftreating a bacterial infection, comprising administering to a subject inneed thereof a therapeutically effective amount of the pharmaceuticalcomposition according to the present subject matter. In an embodiment,the pharmaceutical composition may be organic.

The colostrum or pharmaceutical compositions thereof can be administeredto a subject by any suitable route. For example, the colostrum orpharmaceutical compositions can be administered orally (includingbuccally and sublingually), nasally, rectally, intracisternally,intra-vaginally, intraperitoneally, topically, transdermally (as bypowders, ointments, or drops), and/or parenterally. As used herein,“parenteral” administration refers to modes of administration other thanthrough the gastrointestinal tract, which includes intravenous,intramuscular, intraperitoneal, intrasternal, intramammary, intraocular,retrobulbar, intrapulmonary, intrathecal, subcutaneous andintraarticular injection and infusion. Surgical implantation may also becontemplated, including, for example, embedding a composition of thedisclosure in the body such as, for example, in a tissue, in theabdominal cavity, under the splenic capsule, brain, or in the cornea.

Accordingly, the route of administration can include intranasaladministration, oral administration, inhalation administration,subcutaneous administration, transdermal administration, intradermaladministration, intra-arterial administration with or without occlusion,intracranial administration, intraventricular administration,intravenous administration, buccal administration, intraperitonealadministration, intraocular administration, intramuscularadministration, implantation administration, topical administration,intratumor administration, and/or central venous administration.

The following examples illustrate the present teachings.

Example 1 Determining the Lethal Dose and Optimum Route of Infection forFour Gram-Positive and Gram-Negative Bacteria in a Rat Model

The ethical committee of King Saud University provided prior approvalbefore any animal studies were commenced. All animals were fed withstandard laboratory rat chow (Basal diet 5755, PMI NutritionInternational, Inc., Richmond, Calif., USA) according to the NationalInstitute of Health guidelines. Rats were quarantined for six daysbefore commencing experiments. Chemical restraint anesthesia wasachieved using injectable sedation, such as a Ketamine/Xylazine mixadministered subcutaneously.

The Riyadh Military Hospital Bacteriology Laboratory provided AmericanType Culture Collection (ATCC) Escherichia coli (E. coli) (ATTC 25922).The bacterium is Enteropathogenic E. coli (EPEC) but not EnterotoxigenicE. coli (ETEC). It does not produce verotoxin and is used for mediatesting and as a negative control for LT toxin production. This organismis a CLSI control strain for antimicrobial susceptibility testing. Otherbacterial strains obtained from the same source include Staphylococcusaureus subsp. aureus Rosenbach (S. aureus) (ATCC 29213),Methicillin-resistant Staphylococcus aureus (MRSA) (ATCC 12498),Pseudomonas aeruginosa (P. aeruginosa) (ATCC 27853).

Bacterial propagation was performed in Trypticase Soy Broth (TSB) forseven hours at 26° C., and stock aliquots were frozen at −79° C. untilused. A frozen stock sample was thawed and plated onto Trypticase SoyBlood Agar plates in preparation for a bacterial inoculum lethal dosetrial using rats injected subcutaneously and intravenously. The resultsof this trial are compiled in Table 1 and Table 2 below.

Virus and antibody-free male Wistar rats were acquired from the PharmacyCollege of King Saud University. Animals were housed up to 6 animals ina cage. Male Wistar rats (n=118) weighing 190-300 g were randomlyselected to receive either intravenous (i.v.) or intraperitoneal (i.p.)injection with either E. coli or one of the other three studied bacteria(n=20-40 per group). Control rats were kept in a separate compartment.Animals were observed after bacterial injection to record the number ofdead animals. The results are summarized in Table 1. At six hours afterbacterial administration through either injection route, animalsappeared weak and lethargic. Estimated lethal doses of each pathogenicmicrobe were determined as shown in Table 2. In summary, the optimizedlethal doses were determined to be ˜6×10⁸/ml Colony Forming Units (CFU)for E. coli, 9×10⁸/0.5 ml CFU S. aureus, ˜9.5×10⁸/0.5 ml CFU P.aeruginosa, and 12×10⁸/1.5 ml CFU MRSA. The i.v. route of administrationwas preferred for all microbes.

TABLE 1 Lethal Dose in Rats Demonstration for Studied Groups Over OneWeek Bacterial strains Concentration per ml Injection Site No. Deaths in1 week Escherichia coli 10⁸/0.5 ml i.p. (3) One - 4 days ATTC 2592210⁸/1 ml (3 rats) i.p.(3) i.v.(3) Zero 10⁸/2 ml (3 rats) i.p.(3) i.v.(3)One (i.v.) - 7 days 10⁸/3 ml (6 rats) i.p.(3) Zero 10¹⁶/0.1 ml (2 rats)i.p.(2) Two - 2 days 10¹⁶/0.5 ml (1 rat) i.p.(3) i.v.(3) Zero 10¹⁶/1 ml(1 rat) i.v.(3 + 3) Three - 1 day 10¹⁶/1.5 ml (1 rat) 10¹⁸/1 ml (6 rats)10³⁶/1 ml (6 rats) Pseudomonas aeruginosa 10⁸/0.5 ml (3 rats) i.p.(3)Zero ATCC27853 10⁸/1 ml (2 rats) i.p.(2) i.v.(2) One (i.v., 10⁸/2 ml) 1day 10⁸/2 ml (2 rats) i.p.(1) i.v.(1) Zero 10⁸/3 ml (2 rats)t i.p.(3)One (10⁴⁰/0.5 ml) 3 days 10⁴⁰/0.1 ml (2 rats) i.p.(2) Zero 10⁴⁰/0.5 ml(rat) i.p.(3 + 3) i.v.(3 + 3) Three (i.v., 10⁷²) 3 days 10⁴⁰/1 ml (1rat) 10⁴⁰/1.5 ml (1 rat) 10⁴⁰/0.5 ml (6 rats) 10⁷²/0.5 ml (6 rats)Staphylococcus aureus 10⁸/0.5 ml (3 rats) i.p.(3) One - 4 days ATCC1249810⁸/1 ml (2 rats) i.p.(2) i.v.(2) Zero 10⁸/2 ml (2 rats) i.p.(1) i.v.(1)Zero 10⁸/3 ml (2 rats) i.p.(3) Zero 10²⁰/0.1 ml (2 rats) i.p.(2) Zero10²⁰/0.5 ml (1 rat) i.v.(3 + 3) Three (i.v., 10⁶⁴/0.5 ml) 2 days 10²⁰/1ml (1 rat) 10²⁰/1.5 ml (1 rat) 10⁴⁰/0.5 ml (3 rats) 10⁶⁴/0.5 ml (3 rats)Methicillin-resistant 10⁸/0.5 ml (3 rats) i.p.(3) Zero Staphylococcusaureus 10⁸/1 ml (2 rats) i.p.(2) i.v.(.2) Zero ATCC12498 10⁸/2 ml (2rats) i.p.(1) i.v.(1) Zero 10⁸/3 ml (2 rats) i.p.(3) Zero 410²⁰/0.1 ml(2 rats) i.p.(2) Zero 10²⁰/0.5 ml (1 rat) i.p.(3 + 3) i.v.(3 + 3) Zero10²⁰/1 ml (1 rat) i.v.(3 + 3) Zero 10²⁰/1.5 ml (1 rat) i.p.(3) Seven - 6days 10⁴⁰/0.5 ml (6 rats) i.p.(2) 10⁶⁴/0.5 ml (6 rats) i.v.(2) 10⁶⁰/1 ml(3 rats) 10⁷²/1 ml (3 rats) 10⁹⁰/1 ml (2 rats) 10⁹⁰/1.5 ml (1 rat)10¹²⁰/1 ml (2 rats) 10¹²⁰/1.5 ml (2 rats)

TABLE 2 Recommended Lethal Dose for Studied Bacteria Bacterial strainsConcentration per ml Injection Site No. Deaths in 1 Week Escherichiacoli ~6 × 10⁸/1 ml i.v. (3 rats) Three - 1 day ATTC 25922 CFU/mlPseudomonas aeruginosa ~9.5 × 10⁸/0.5 ml i.v. (3 rats) Three - 3 daysATCC27853 CFU/ml Staphylococcus aureus 9 × 10⁸/0.5 ml i.v. (3 rats)Three - 2 days ATCC12498 CFU/ml Methicillin-resistant 12 × 10⁸/1.5 mli.v. (3 rats) Three - 6 days Staphylococcus aureus CFU/ml ATCC12498

Example 2 Determining Effect of Camel Colostrum, Bovine Colostrum, and aMixture of Camel Colostrum and Bovine Colostrum on Survival of RatsInfected with Gram-Positive and Gram-Negative Bacteria

Three types of colostrum were used from different sources. Camelcolostrum was sourced from 10 healthy individual camels (Camelusdromedarius) that were chosen randomly (average age, six years) from theConservation and Genetic Improvement Center in Al-karj, Saudi Arabia.Samples were obtained manually from all camels' udders and werecollected immediately after parturition.

Bovine colostrum samples were sourced from 10 healthy Holstein cows(average age, 3.8 years) provided by the Almarai Company. The ImmunologyLab, King Khaled Hospital, Medicine College, King Saud UniversityHospital, collected the samples and confirmed that the source cows werein good health and had no clinical evidence of mastitis or tuberculosis,delivered healthy full-term infants, and had not consumed medicationswithin one week before collection.

The colostrum samples were collected using the following protocol: 20samples collected at the first-day parturition after thorough handwashing and cleansing of the breast and nipple with soap and tap water.Milk samples were expressed manually into sterile Erlenmeyer flasks,pooled, dispensed into sterile test tubes and stored on at −20° C.

The study included a control group with no graft contamination and noantibiotic prophylaxis (Group 1), a bacterial treatment group (Group 2),a colostrum treatment group (Group 3), and a bacterial treatment andcolostrum treatment group (Group 4). (See Table 3) Group 3 includedthree different sub-groups, designated camel colostrum, bovinecolostrum, and mix colostrum. One ml of prepared colostrum solution (28g lyophilized colostrum/liter sterilized saline solution (w/v)) wasadministered by intraperitoneal injection every day early in the morningfor 30 days.

The rats of Group 2 and Group 4 were then anesthetized with chloroformor ether and injected intravenously with the recommended lethal dose ofone of the studied bacteria in the tail vein. Survival rates in eachgroup were observed for two weeks after injection.

TABLE 3 The Study Groups Group 1 Control group: Sterilized saline wasadministered every day at eight AM for 30 days Group 2 Bacterial groups:four bacterial sub-groups were designated, one for each bacterium:Escherichia coli ATTC 25922 Pseudomonas aeruginousa ATCC27853Staphylococcus aureus ATCC12498 Methicillin-resistant Staphylococcusaureus ATCC12498 The lethal dose of each bacterium established in Table1 was administered intravenously once in the tail vein of each rat usinga 14-gauge needle at eight AM on the starting day. Group 3 Colostrumgroups: Three colostrum sub-groups were designated: Camel ColostrumBovine Colostrum Mix Colostrum One ml of prepared colostrum solution (28g lyophilized colostrum/liter sterilized saline solution (w/v)) wasinjected i.p. daily at eight AM for 30 days. Group 4 Bacterial andColostrum groups: Sub-groups were designated for all possiblecombinations of Group 2 and Group 3.

At zero time up to one month and after inoculation with the four studiedbacteria and induction by colostrum as in Table 4 and Table 5, animalsappeared weak and lethargic with the microbial treatments. In controlswithout bacterial treatment, camel colostrum, bovine colostrum, and mix(a mixture of both camel and bovine colostrum) produced a similarsurvival rate without evidence of concentration of disease (See Table4). Compared with the control treatment, bacterial treatment withcolostrum improved mortality rates associated with the bacterialvariable and route of treatment. These animals lived through one monthof observation. The death of animals started from the second day untilthe 22^(nd) day, with most deaths occurring in the administratedpathogenic bacteria only group (Group 2). As shown in Table 4, thefastest and the best-protected treatment was lethal S. aureus with50-100% mortality.

Moreover, 100% of rats injected with S. aureus and vaccinated i.p. byone of the three types of colostrum survived. Camel colostrum had atremendous protective effect from lethal MRSA, converting 100% mortalityto 100% survival. Camel and bovine colostrum enhanced survival againstlethal P. aeruginosa from 0% to 33% survival in the former and from 50%to 66% survival in the latter (see Table 4). Lethal E. coli caused 84%mortality, and camel colostrum enhanced survival of up to 83%. On theother hand, mixed colostrum also enhanced survival, up to 50%.

TABLE 4 Morbidity and Mortality (Survival %) in Lethal Dose Inductionfor Four Pathogenic Bacteria using Three Colostrum Groups asAnti-Microbial Agents Colostrum Bacteria Healthy Bacteria & ColostrumHealthy Dose Rats per Healthy Healthy Rats or Dead(Day) Rats orDead(Day) Rats (~CFU/ml) Group Rats Camel Bovine Mix Camel Bovine MixCamel Bovine Mix E. coli: 6 6 6 6 6 6 2(3) 6 1(2) 1(2)  3(22) 6 × 10⁸/ml2(7) 2(9)   1(13) 3(16) % Survival 100 100 100 100 100 100  16 100 83 050 P. aeruginosa: 6 6 6 6 6 2(2) 1(2) 6 2(5) 2(20) 6 9.5 × 10⁸/0.5 ml3(3) 1(8) 1(9) 1(7)  1(15)  1(11) % Survival 100 100 100 100 100 0 50100 33 66 100 S. aureus 6 6 6 6 6 1(3) 2(4) 1(3) 6 6 6 9 × 10⁸/0.5 ml1(5) 2(7) 1(4)  1(17)  1(10)  1(12)  1(11) % Survival 100 100 100 100100 50   0 50 100 100 100 MRSA: 6 6 6 6 6 2(2)  6 6 6 6 6 12 × 10⁸/1.5ml 1(4) 1(8)  1(12)  1(16) % Survival 100 100 100 100 100 0 100  100 100100 100

Example 3 Determining Effect of Camel Colostrum, Bovine Colostrum, and aMixture of Camel Colostrum and Bovine Colostrum on Percentage Change inWeight of Rats Infected with Gram-Positive and Gram-Negative Bacteria

The effect of the treatment on the percentage change of the weight ofrats (grams) for each studied pathogenic bacterium was determined usingthree types of colostrum; Camel, Bovine, and Mix. The results of thisexperiment are illustrated in Table 5 and Table 6. The range of percentchange in the weight of healthy rats was revealed in minimum and maximum(Min 10%-Max 29%) through one month of observation. Camel colostrum andMix colostrum induced a remarkable 6%-59% increase in the weight ofrats. Bacterial injections dramatically decreased the percent weightchange in rats from −12% to 12%. The most notable max increase values inpercent change of weight of rats after colostrum treatment were observedwhen comparing the weight of rats infected with MRSA alone with theweight of the rats treated with mix colostrum. Mix colostrum induced 43%change in the weight of rats compared to −21% weight change in ratsinjected with a lethal dose of MRSA alone.

TABLE 5 Treatment Effects on Average % Change in Weight (g) of Rats(Healthy & Bacteria Treatment Groups) Group Healthy Bacteria Day 1 10 2030 Avg % 1 10 20 30 Avg % Camel Colostrum E. coli 226 256 276 292 29%210 205 209 214 1.9 P. aeruginosa 236 215 250 287 21% 228 232 230 2457.4 S. aureus 227 205 228 250 10% 230 239 243 235 2.1 MRSA 235 225 247290 23% 207 210 222 205 −0.9 Bovine Colostrum E. coli 226 256 276 29229% 215 224 230 235 9.3 P. aeruginosa 236 215 250 287 21% 234 230 227220 −5.9 S. aureus 251 205 228 250 10% 215 224 230 235 9.3 MRSA 235 225247 290 23% 234 230 227 220 −5.9 Mix Colostrum E. coli 226 256 276 29229% 242 250 267 259 7 P. aeruginosa 236 215 250 287 21% 247 255 247 27912 S. aureus 251 205 228 250 10% 273 249 231 213 −21 MRSA 235 225 247290 23% 252 270 263 267 5.9

TABLE 6 Treatment Effects on Average % Change in Weight (g) of Rats(Bacteria & Colostrum and Colostrum Treatment Groups) Group Bacteria &Colostrum Colostrum Day 1 10 20 30 Avg % 1 10 20 30 Avg % CamelColostrum E. coli 227 235 245 240 5.7%  213 246 278 286 34 P. aeruginosa260 252 254 259 −0.3%  227 239 345 354 55 S. aureus 234 239 244 260 11%243 275 288 295 21 MRSA 250 253 255 259 3.6%  252 258 261 269 6 BovineColostrum E. coli 236 264 285 298 26% 230 236 254 270 17 P. aeruginosa241 265 273 295 37% 242 265 277 293 21 S. aureus 236 264 285 298 26% 230236 254 270 17 MRSA 241 265 273 295 22.4 218 244 279 298 36 MixColostrum E. coli 270 284 312 326 20% 205 240 276 288 40 P. aeruginosa234 255 288 310 32% 272 281 301 321 18 S. aureus 230 252 284 329 43% 209225 246 280 33 MRSA 202 235 272 280 38% 193 225 286 307 59

Example 4

Effect of Infection and/or Colostrum on Expression of Three Cytokines inRats

For circulating blood analysis, animals were anesthetized with an ethermask. Circulating levels of IFN-γ, IL-10, and TNF-α were measured inserum samples of all rats obtained from the eye vein. Blood samples werecollected in heparinized syringes to asses cytokine level assessed ofIFN-γ, IL10, and TNF-α. Serum concentrations of each cytokine weremonitored at 0 days post-treatment, 24 hours post-treatment, 7-25 dayspost-treatment (midpoint or Mid), and 30 days post-treatment (Last orFinal) by ELISA. Briefly, blood samples (2 ml) were centrifuged for 10minutes at 3500 rpm, at 4° C., and the serum samples were kept at −20°C. in a freezer until examined. Commercial reagents used to identifyindividual cytokines included Quantikine®ELISA, Rat TNF-α Immunoassay,Catalog Number RTA00, Quantikine®ELISA, Rat IL-10 Immunoassay,Quantikine®ELISA, and Rat IFN-γ Immunoassay. These kits were usedaccording to the manufacturer's recommendations (USA & Canada R&DSystems, Inc. Minneapolis). The limits of sensitivity of the assay weredetermined to be five pg/ml for TNF-α and ten pg per ml for IFN-γ andIL-10, respectively. The intra- and inter-assay coefficients ofdeviation were less than 5% and 10%, respectively.

Quantitative data were statistically represented in terms of minimum,maximum, and median. A comparison between different groups in thepresent study was done using the Kruskal-Wallis test to compare betweenmore than two nonparametric groups.

Correlation between various variables was determined using Spearman rankcorrelation coefficient (R) with graph representations using linearregression.

A probability value (p-value) less than or equal to 0.05 was consideredsignificant. All statistical analyses were performed using statisticalsoftware SPSS (Statistical Package for Social Science) statisticalprogram version 16.0. Graphs were created using SPSS statistical programversion 16.0 with Microsoft Excel program version 2010.

Initial results for IFN-γ expression as a measure of the immune responsein the presence of colostrum, E. coli, or E. coli and colostrum arepresented in Table 7 and FIG. 1.

TABLE 7 E. Coli and IFN-y Immune Response in the Presence of Colostrum(Expressed as Median pg/ml (Min-Max)) Colostrum Type Camel ColostrumBovine Colostrum Mix Colostrum P value^(A) Healthy Control  2.20(0.00-43.91) 2.20 (0.00-43.91)  2.20 (0.00-43.91) 1.000 Colostrum 0.00(0.00-0.03) 0.09 (0.00-209.50) 17.82 (0.00-125.00) 0.055 Bacteria 0.02(0.00-0.07) 0.06 (0.01-74.24)   0.11 (0.00-125.00) 0.219 Colostrum &0.01 (0.00-0.03) 0.06 (0.00-117.10) 0.12 (0.00-93.75) 0.098 Bacteria Pvalue^(B) 0.018 0.808 0.901 ^(A)= Comparison between differenttreatments within a single colostrum type using a nonparametric test(Kruskal-Wallis Test). ^(B)= Comparison between different colostrumtypes in each treatment using a nonparametric test (Kruskal-WallisTest).

Data from the full month-long experiment are presented in Table 8 andFIGS. 2A-2C. In the model of E. coli bacterial infection treated witheither bovine colostrum or with the mixed colostrum, IFN-γ elevated upto 0.12 pg/ml in the mixed colostrum treatment group but was 0.06 pg/mlin the bovine colostrum treatment group. At the end of the trial (Last)IFN-γ exhibited a steady level of the immune response in the mixedcolostrum treatment group (93.750 pg/ml) as well as the bacterialinfection and mixed colostrum treatment group. (See Table 8). Thisexperiment confirms the synergistic effect of mixed colostrum, as ittriggers a more significant immune response than bovine colostrum orcamel colostrum alone.

TABLE 8 E. coli and IFN-γ Immune Response in the Presence of Colostrumat 0 h, 24 h, 7-25 days (M), and 30 days (Last) (Expressed as Median(Min-Max) Treatment Group Time Camel Bovine Mix Healthy 0 h1.496(0.220-2.770) 1.496(0.220-2.770) 1.496(0.220-2.770) 24 h3.587(1.630-5.550) 3.587(1.630-5.550) 3.587(1.630-5.550) M 42.175(40.440-43.910)  42.175(40.440-43.910)  42.175(40.440-43.910)Last 0.005(0.000-0.010) 0.005(0.000-0.010) 0.005(0.000-0.010) P value0.104 0.104 0.104 Colostrum 0 h 0.019(0.010-0.030) 1.055(0.120-1.990)3.129(0.000-6.260) 24 h 0.002(0.000-0.000) 0.058(0.060-0.060)1.667(0.000-3.330) M 0.002(0.000-0.000) 128.800(48.10-209.50)  30.327(29.390-31.260) Last 0.005(0.000-0.010) 0.001(0.000-0.000)93.750(62.50-125.00) P value 0.160 0.078 0.106 Bacteria 0 h0.025(0.010-0.040) 1.019(0.050-1.990) 0.003(0.000-0.000) 24 h0.003(0.000-0.010) 0.051(0.040-0.070) 0.003(0.000-0.000) M0.016(0.000-0.030) 37.183(0.130-74.240) 19.633(0.200-39.060) Last0.049(0.020-0.070) 0.013(0.010-0.020)  125.00(125.00-125.00) P value0.280 0.139 0.103 Colostrum & 0 h 0.018(0.010-0.030) 0.048(0.020-0.070)0.061(0.000-0.120) Bacteria 24 h 0.001(0.000-0.000) 0.067(0.040-0.090)0.008(0.010-0.010) M 0.016(0.000-0.030) 77.905(38.71-117.10) 20.840(20.840-20.840) Last 0.000(0.000-0.000) 0.012(0.000-0.020) 93.750(93.750-93.750) P value 0.165 0.104 0.362

In the model of E. coli bacterial infection, both camel colostrum andmixed colostrum stimulated TNF-α. The highest enhancement was for camelcolostrum in all treatments (0.56 pg/ml, 0.57 pg/ml, and 0.60 pg/ml forcolostrum alone, bacteria alone, and colostrum with bacteria,respectively). These results are summarized in Table 9 and FIG. 3. Datafrom the full month-long experiment are presented in FIGS. 4A-4C.

TABLE 9 E. coli and TNF-α Immune Response in the Presence of Colostrum(Expressed as Median pg/ml (Min-Max)) Colostrum Type Camel Bovine Mix Pvalue^(A) Healthy 0.00 (0.00-0.60) 0.00 (0.00-0.60) 0.00 (0.00-0.60)1.000 Colostrum 0.56 (0.00-0.94)  0.00 (0.00-194.80) 0.03 (0.00-0.62)0.261 Bacteria 0.57 (0.00-1.13) 0.00 (0.00-0.22) 0.18 (0.00-0.96) 0.033Colostrum & 0.60 (0.25-1.00) 0.00 (0.00-0.00) 0.23 (0.00-0.76) 0.002Bacteria P value^(B) 0.101 0.331 0.373 ^(A)= Comparison betweendifferent treatments within a single Colostrum type using aNonparametric Test (Kruskal-Wallis Test). ^(B)= Comparison betweendifferent Colostrum types in each treatment using a Nonparametric Test(Kruskal-Wallis Test).

In the model of E. coli bacterial infection, mixed colostrum alonetriggered the IL-10 immune response more than the other treatments(83.30 pg/ml). The most enormous increase was observed in the presenceof mixed colostrum and E. coli bacteria (1000.0 pg/ml at the Lastpoint). (See Table 10 and FIG. 5)

TABLE 10 E. coli and IL-10 Immune Response in the Presence of Colostrum(Expressed as Median pg/ml (Min-Max)) Colostrum Type Camel Bovine Mix Pvalue^(A) Healthy 0.00 (0.00-36.76) 0.00 (0.00-36.76) 0.00 (0.00-36.76) 1.000 Colostrum 0.00 (0.00-16.30) 0.14 (0.00-38.65) 83.30 (0.00-1000.00)0.115 Bacteria 0.00 (0.00-0.64)  0.02 (0.00-0.26)  31.62 (0.00-1000.00)0.159 Colostrum & 0.00 (0.00-0.14)  0.01 (0.00-0.29)   0.01(0.00-1000.00) 0.290 Bacteria P value^(B) 0.750 0.353 0.632 ^(A)=Comparison between different treatments within a single Colostrum typeusing a Nonparametric Test (Kruskal-Wallis Test). ^(B)= Comparisonbetween different Colostrum types in each treatment using aNonparametric Test (Kruskal-Wallis Test).

Data from the full month-long experiment are presented in Table 11 andFIGS. 6A-6C. IL-10 started to increase after 24 hours in the bacterialinfection with 80.308 pg/ml. Moreover, colostrum continued to induceIL-10 from 7 days-25 days up to 166.60 pg/ml.

TABLE 11 E. coli and IL-10 Immune Response in the Presence of Colostrumat 0 h, 24 h, 7-25 days (M), and 30 days (Last) (Expressed as Median(Min-Max)) Treatment Group Time Camel Bovine Mix Healthy 0 h 0.016(0.000-0.030) 0.016 (0.000-0.030) 0.016 (0.000-0.030) 24 h 18.378(0.000-36.760) 18.378 (0.000-36.760) 18.378 (0.000-36.760) M 0.018(0.000-0.030) 0.018 (0.000-0.030) 0.018 (0.000-0.030) Last 0.004(0.000-0.010) 0.004 (0.000-0.010) 0.004 (0.000-0.010) P value 0.9350.935 0.935 Colostrum 0 h 0.000 (0.000-0.000) 19.621 (0.590-38.650)0.001 (0.000-0.000) 24 h 0.000 (0.000-0.000) 0.810 (0.150-1.470) 0.002(0.000-0.000) M  8.152 (0.000-16.300) 0.077 (0.020-0.130) 166.63(166.60-166.66) Last 0.005 (0.000-0.000) 0.001 (0.000-0.000) 750.0(500.0-1000.0) P value 0.105 0.104 0.091 Bacteria 0 h 0.000(0.000-0.000) 0.156 (0.050-0.260) 0.010 (0.000-0.020) 24 h 0.318(0.000-0.640) 0.017 (0.010-0.020) 80.308 (63.220-97.400) M 0.009(0.000-0.020) 0.044 (0.020-0.070) 0.001 (0.000-0.000) Last 0.056(0.060-0.060) 0.002 (0.000-0.000) 750.0 (500.0-1000.0) P value 0.4670.139 0.080 Colostrum & 0 h 0.000 (0.000-0.000) 0.162 (0.030-0.290)0.005 (0.000-0.010) Bacteria 24 h 0.000 (0.000-0.000) 0.010(0.010-0.010) 0.013 (0.010-0.010) M 0.006 (0.000-0.010) 0.010(0.010-0.010) 0.001 (0.000-0.000) Last 0.089 (0.040-0.140) 0.006(0.000-0.010) 1000.0 (1000.0-1000.0) P value 0.116 0.276 0.284

In the model of P. aeruginosa infection, the mixed colostrum inducedIFN-γ expression of 17.82 pg/ml. Further, the mixed colostrum combinedwith bacterial infection resulted in an IFN-γ expression of 32.22 pg/ml.These results are presented in Table 12 and FIG. 7. Data from the fullmonth-long experiment are presented in FIGS. 8A-8C.

TABLE 12 P. aeruginosa and IFN-γ Immune Response in the Presence ofColostrum (Expressed as Median pg/ml (Min-Max)) Colostrum Type CamelBovine Mix P value^(A) Healthy 2.20 (0.00-43.91) 2.20 (0.00-43.91)  2.20(0.00-43.91) 1.000 Colostrum 0.00 (0.00-0.03)  0.09 (0.00-209.50) 17.82(0.00-125.00) 0.055 Bacteria 0.01 (0.00-74.40) 0.07 (0.02-105.79) 0.50(0.01-62.50) 0.284 Colostrum 0.00 (0.00-0.00)  0.06 (0.03-0.49)  32.22(0.00-457.20) 0.002 &Bacteria P value^(B) 0.007 0.712 0.578 ^(A)=Comparison between different treatments within a single Colostrum typeusing a Nonparametric Test (Kruskal-Wallis Test). ^(B)= Comparisonbetween different Colostrum types in each treatment using aNonparametric Test (Kruskal-Wallis Test).

In the model of P. aeruginosa infection, the most considerableenhancement of TNF-α expression was detected in the group administeredcamel colostrum, which demonstrated 0.56 pg/ml, 1.54 pg/ml and 1.00pg/ml for colostrum treatment alone, bacterial treatment alone, andcolostrum and bacterial treatment, respectively. These results arepresented in Table 13 and FIG. 9.

TABLE 13 P. aeruginosa and TNF-α Immune Response in the Presence ofColostrum (Expressed as Median pg/ml (Min-Max)) Colostrum Type CamelBovine Mix P value^(A) Healthy 0.00 (0.00-0.60) 0.00 (0.00-0.60) 0.00(0.00-0.60) 1.000 Colostrum 0.56 (0.00-0.94)  0.00 (0.00-194.80) 0.03(0.00-0.62) 0.261 Bacteria 1.54 (0.54-1.91) 0.00 (0.00-0.00) 0.00(0.00-1.09) 0.004 Colostrum&  1.00 (0.00-12.50) 0.00 (0.00-0.00) 0.55(0.00-4.58) 0.006 Bacteria P value^(B) 0.025 0.185 0.663 ^(A)=Comparison between different treatments within a single Colostrum typeusing a Nonparametric Test (Kruskal-Wallis Test). ^(B)= Comparisonbetween different Colostrum types in each treatment using aNonparametric Test (Kruskal-Wallis Test).

Data from the month-long experiment are presented in Table 14 and FIGS.10A-10C. P. aeruginosa administration increased TNF-α expression after24 hours to 1.171 pg/ml. In comparison, an expression level of 0.828pg/ml was detected for camel colostrum administration. The mostefficient change in TNF-α expression was observed for the administrationof both camel colostrum and P. aeruginosa (8.931 pg/ml 24 hours afteradministration of the bacterium).

TABLE 14 P. aeruginosa and TNF-α Immune Response in the Presence ofColostrum at 0 h, 24 h, 7-25 days (M), and 30 days (Last) (Expressed asMedian(Min-Max)) Treatment Time Camel Bovine Mix Healthy 0 h0.300(0.000-0.600) 0.300(0.000-0.600) 0.300(0.000-0.600) 24 h0.220(0.000-0.440) 0.220(0.000-0.440) 0.220(0.000-0.440) M0.269(0.000-0.540) 0.269(0.000-0.540) 0.269(0.000-0.540) Last0.000(0.000-0.000) 0.000(0.000-0.000) 0.000(0.000-0.000) P value 0.6750.675 0.675 Colostrum 0 h 0.558(0.540-0.580) 0.000(0.000-0.000)0.000(0.000-0.000) 24 h 0.828(0.720-0.940) 0.000(0.000-0.000)0.000(0.000-0.000) M 0.430(0.170-0.690) 104.749(14.70-194.80) 0.177(0.050-0.300) Last 0.000(0.000-0.000) 0.000(0.000-0.000)0.490(0.360-0.620) P value 0.108 0.077 0.078 Bacteria 0 h1.594(1.280-1.910) 0.000(0.000-0.000) 0.000(0.000-0.000) 24 h1.171(0.540-1.800) 0.000(0.000-0.000) 0.000(0.000-0.000) M —0.000(0.000-0.000) 0.185(0.000-0.360) Last — 0.000(0.000-0.000)0.550(0.010-1.090) P value 0.439 1.000 0.100 Colostrum& 0 h0.769(0.540-1.000) 0.000(0.000-0.000) 0.000(0.000-0.000) Bacteria 24 h 8.931(5.360-12.500) 0.000(0.000-0.000) 0.000(0.000-0.000) M0.861(0.360-1.360) 0.000(0.000-0.000) 2.568(0.550-4.580) Last0.000(0.000-0.000) 0.000(0.000-0.000) 0.695(0.560-0.830) P value 0.1851.000 0.183

In the model of P. aeruginosa infection, the administration of themixture of colostrum alone caused the greatest change in IL-10 levels(83.30 pg/ml), as shown in Table 15 and FIG. 11. Data from themonth-long experiment are presented in Table 16 and FIGS. 12A-12C.

TABLE 15 P. aeruginosa and IL-10 Immune Response in the Presence ofColostrum (Expressed as Median pg/ml (Min-Max)) Colostrum Type CamelBovine Mix P value^(A) Healthy 0.00(0.00-36.76) 0.00(0.00-36.76)0.00(0.00-36.76)  1.000 Colostrum 0.00(0.00-16.30) 0.14(0.00-38.65)83.30(0.00-1000.00) 0.115 Bacteria 0.00(0.00-0.01)  0.01(0.01-0.03) 0.07(0.00-500.00) 0.019 Colostrum & 0.00(0.00-72.69) 0.08(0.00-1.67) 1.31(0.00-500.00) 0.388 Bacteria P value^(B) 0.749 0.321 0.286 ^(A)=Comparison between different treatments within a single Colostrum typeusing a Nonparametric Test (Kruskal-Wallis Test). ^(B)= Comparisonbetween different Colostrum types in each treatment using aNonparametric Test (Kruskal-Wallis Test).

TABLE 16 P. aeruginosa and IL-10 Immune Response in the Presence ofColostrum at 0 h, 24 h, 7-25 days (M), and 30 days (Last) (Expressed asMedian(Min-Max)) Treatment Time Camel Bovine Mix Healthy 0 h0.016(0.000-0.030) 0.016(0.000-0.030) 0.016(0.000-0.030) 24 h18.378(0.000-36.760) 18.378(0.000-36.760) 18.378(0.000-36.760) M0.018(0.000-0.030) 0.018(0.000-0.030) 0.018(0.000-0.030) Last0.004(0.000-0.010) 0.004(0.000-0.008) 0.004(0.000-0.010) P value 0.9350.935 0.935 Colostrum 0 h 0.000(0.000-0.000) 19.621(0.590-38.650)0.001(0.000-0.000) 24 h 0.000(0.000-0.000) 0.810(0.150-1.470)0.002(0.000-0.000) M  8.152(0.000-16.300) 0.077(0.020-0.130) 166.63(166.60-166.66) Last 0.005(0.000-0.000) 0.001(0.001-0.001)750.00(500.0-1000.0) P value 0.105 0.104 0.091 Bacteria 0 h0.001(0.001-0.001) 0.020(0.010-0.030) 0.029(0.020-0.040) 24 h0.004(0.000-0.008) 0.007(0.010-0.010)  8.914(0.070-17.760) M —0.008(0.010-0.010)  83.301(0.000-166.600) Last — 0.009(0.010-0.010) 500.00(500.00-500.00) P value 1.000 0.701 0.375 Colostrum& 0 h0.000(0.000-0.000) 0.008(0.000-0.020) 0.043(0.040-0.050) Bacteria 24 h36.414(0.140-72.690) 1.173(0.670-1.670) 3.653(2.570-4.740) M3.601(0.000-7.200) 0.030(0.030-0.030) 0.002(0.001-0.002) Last0.002(0.000-0.000) 0.128(0.130-0.130)  500.00(500.00-500.00) P value0.297 0.194 0.080

In the model of S. aureus subsp. aureus Rosenbach infection, IFN-γlevels increased to 78.10 pg/ml when the mixed colostrum and bacteriawere administered, as compared to 17.82 pg/ml when the mixed colostrumalone was administered (see Table 17 and FIG. 13).

TABLE 17 S. aureus and IFN-γ Immune Response in the Presence ofColostrum (Expressed as Median pg/ml (Min-Max)) Colostrum Type: CamelBovine Mix P value^(A) Healthy 2.20(0.00-43.91) 2.20(0.00-43.91)2.20(0.00-43.91) 1.000 Colostrum 0.00(0.00-0.03)   0.09(0.00-209.50)17.82(0.00-125.00) 0.055 Bacteria 0.01(0.00-64.78) 0.44(0.04-75.23)0.50(0.01-62.50) 0.076 Colostrum & 0.02(0.00-2.91)  23.76(0.01-94.87)  78.10(0.03-6541.00) 0.008 Bacteria P value^(B) 0.026 0.436 0.362 ^(A)=Comparison between different treatments within a single Colostrum typeusing a Nonparametric Test (Kruskal-Wallis Test). ^(B)= Comparisonbetween different Colostrum types in each treatment using aNonparametric Test (Kruskal-Wallis Test).

In the month-long experiment, administration of the mixture of colostrumand S. aureus resulted in an increased level of IFN-γ of 3333.0 pg/ml bythe final time-point. In comparison, at days 7-25 (M), the median IFN-γlevel of rats administered mixed colostrum and S. aureus was 278.15pg/ml. At the 24 hour time-point this level was only 15.647 pg/ml. Theseresults are presented in Table 18 and FIGS. 14A-14C.

TABLE 18 S. aureus and IFN-γ Immune Response in the Presence ofColostrum at 0 h, 24 h, 7-25 days (M), and 30 days (Last) (Expressed asMedian(Min-Max)) Treatment Time Camel Bovine Mix Healthy 0 h1.496(0.220-2.770) 1.496(0.220-2.770) 1.496(0.220-2.770) 24 h3.587(1.630-5.550) 3.587(1.630-5.550) 3.587(1.630-5.550) M 42.175(40.440-43.910)  42.175(40.440-43.910)  42.175(40.440-43.910)Last 0.005(0.000-0.010) 0.005(0.000-0.010) 0.005(0.000-0.010) P value0.104 0.104 0.104 Colostrum 0 h 0.019(0.008-0.030) 1.055(0.120-1.990)3.129(0.000-6.260) 24 h 0.002(0.000-0.004) 0.058(0.060-0.060)1.667(0.000-3.330) M 0.002(0.001-0.002) 128.800(48.10-209.50)  30.327(29.390-31.260) Last 0.005(0.003-0.006) 0.001(0.001-0.001)93.750(62.50-125.00) P value 0.160 0.078 0.106 Bacteria 0 h32.400(0.020-64.780)  6.787(0.090-13.480) 0.194(0.110-0.280) 24 h0.066(0.001-0.130) 0.356(0.050-0.670) 2.022(0.710-3.330) M0.005(0.003-0.007) 2.241(0.210-4.270) 10.450(0.070-20.830) Last0.002(0.000-0.002) 37.639(0.040-75.230) 31.256(0.010-62.500) P value0.426 0.919 0.881 Colostrum & 0 h 1.558(0.210-2.910)47.440(0.010-94.870) 0.140(0.030-0.250) Bacteria 24 h 0.010(0.000-0.010)0.355(0.360-0.360) 15.647(0.090-31.200) M 0.008(0.000-0.010) 18.174(12.590-23.760)  278.15(173.90-382.40) Last 0.033(0.020-0.050) 46.956(37.680-56.230) 3333.00(125.0-6541.0)  P value 0.106 0.587 0.139

In the model of S. aureus subsp. aureus Rosenbach infection, the mostexcellent enhancement in TNF-α levels was observed after administrationof camel colostrum, which had 0.56 pg./m., 1.21 pg./m and 0.50 pg./m forColostrum, Bacteria, and Colostrum with Bacteria respectively (See Table19 and FIG. 15). Data from the month-long experiment are presented inTable 20 and FIGS. 16A-16C.

TABLE 19 S. aureus and TNF-α Immune Response in the Presence ofColostrum (expressed as median pg/ml (Min-Max)) Colostrum Type: CamelBovine Mix P value^(A) Healthy 0.00(0.00-0.60) 0.00(0.00-0.60)0.00(0.00-0.60) 1.000 Colostrum 0.56(0.00-0.94)  0.00(0.00-194.80)0.03(0.00-0.62) 0.261 Bacteria 1.21(0.00-9.85) 0.00(0.00-0.00)0.00(0.00-1.09) 0.002 Colostrum & 0.50(0.00-6.55) 0.00(0.00-0.00) 0.12(0.00-116.44) 0.014 Bacteria P value^(B) 0.043 0.118 0.931 ^(A)=Comparison between different treatments within a single Colostrum typeusing a Nonparametric Test (Kruskal-Wallis Test). ^(B)= Comparisonbetween different Colostrum types in each treatment using aNonparametric Test (Kruskal-Wallis Test).

TABLE 20 S. aureus and TNF-α Immune Response in the Presence ofColostrum at 0 h, 24 h, 7-25 days (M), and 30 days (Last) (Expressed asMedian(Min-Max)) Treatment Time Camel Bovine Mix Healthy 0 h0.300(0.000-0.600) 0.300(0.000-0.600) 0.300(0.000-0.600) 24 h0.220(0.000-0.440) 0.220(0.000-0.440) 0.220(0.000-0.440) M0.269(0.000-0.540) 0.269(0.000-0.540) 0.269(0.000-0.540) Last0.000(0.000-0.000) 0.000(0.000-0.000) 0.000(0.000-0.000) P value 0.6750.675 0.675 Colostrum 0 h 0.558(0.540-0.580) 0.000(0.000-0.000)0.000(0.000-0.000) 24 h 0.828(0.720-0.940) 0.000(0.000-0.000)0.000(0.000-0.000) M 0.430(0.170-0.690) 104.749(14.70-194.80) 0.177(0.050-0.300) Last 0.000(0.000-0.000) 0.000(0.000-0.000)0.490(0.360-0.620) P value 0.108 0.077 0.078 Bacteria 0 h5.930(2.010-9.850) 0.000(0.000-0.000) 0.000(0.000-0.000) 24 h0.962(0.720-1.210) 0.000(0.000-0.000) 0.000(0.000-0.000) M0.900(0.580-1.220) 0.000(0.000-0.000) 0.185(0.000-0.360) Last0.000(0.000-0.000) 0.000(0.000-0.000) 0.550(0.010-1.090) P value 0.1851.000 0.100 Colostrum & 0 h 6.184(5.820-6.550) 0.000(0.000-0.000)0.000(0.000-0.000) Bacteria 24 h 0.600(0.580-0.620) 0.000(0.000-0.000)0.000(0.000-0.000) M 0.352(0.280-0.430) 0.000(0.000-0.000) 58.336(0.230-116.440) Last 0.000(0.000-0.000) 0.000(0.000-0.000)0.589(0.560-0.620) P value 0.080 1.000 0.109

In the model of S. aureus subsp. aureus Rosenbach infection, treatmentwith mixed colostrum alone and treatment with mixed colostrum andbacteria led to the highest increase in IL-10 levels (See Table 21 andFIG. 17).

TABLE 21 S. aureus and IL-10 Immune Response in the Presence ofColostrum (Expressed as Median pg/ml (Min-Max)) Colostrum Type: CamelBovine Mix P value^(A) Healthy 0.00(0.00-36.76) 0.00(0.00-36.76)0.00(0.00-36.76)  1.000 Colostrum 0.00(0.00-16.30) 0.14(0.00-38.65)83.30(0.00-1000.00) 0.115 Bacteria 0.00(0.00-0.98)  0.04(0.00-23.78)0.07(0.00-500.00) 0.023 Colostrum & 0.00(0.00-0.07)  0.03(0.00-0.39) 83.38(0.01-1000.00) 0.021 Bacteria P value^(B) 0.677 0.428 0.153 ^(A)=Comparison between different treatments within a single Colostrum typeusing a Nonparametric Test (Kruskal-Wallis Test). ^(B)= Comparisonbetween different Colostrum types in each treatment using aNonparametric Test (Kruskal-Wallis Test).

In the month-long experiment, treatment with mixed colostrum andtreatment with either mixed colostrum alone or with mixed colostrum andbacteria resulted in a similar increase in IL-10 levels (166.63 and166.60 respectively at 7-25 days and 750.00 pg/ml at the end of themonth). These results are presented in Table 22 and FIGS. 18A-18C.

TABLE 22 S. aureus and IL-10 Immune Response in the Presence ofColostrum at 0 h, 24 h, 7-25 days (M), and 30 days (Last) (Expressed asMedian(Min-Max)) Treatment Time Camel Bovine Mix Healthy 0 h0.016(0.000-0.030) 0.016(0.000-0.030) 0.016(0.000-0.030) 24 h18.378(0.000-36.760) 18.378(0.000-36.760) 18.378(0.000-36.760) M0.018(0.000-0.030) 0.018(0.000-0.030) 0.018(0.000-0.030) Last0.004(0.000-0.010) 0.004(0.000-0.010) 0.004(0.000-0.010) P value 0.9350.935 0.935 Colostrum 0 h 0.000(0.000-0.000) 19.621(0.590-38.650)0.001(0.001-0.001) 24 h 0.000(0.000-0.000) 0.810(0.150-1.470)0.002(0.001-0.002) M  8.152(0.000-16.300) 0.077(0.020-0.130) 166.63(166.60-166.66) Last 0.005(0.005-0.005) 0.001(0.000-0.001)750.00(500.0-1000.0) P value 0.105 0.104 0.091 Bacteria 0 h0.491(0.000-0.980) 0.089(0.010-0.170) 0.029(0.020-0.040) 24 h0.000(0.000-0.000) 0.040(0.020-0.060)  8.914(0.070-17.760) M0.000(0.000-0.000) 11.890(0.000-23.780)  83.301(0.000-166.600) Last0.009(0.010-0.010) 0.103(0.010-0.200)  500.00(500.00-500.00) P value0.166 0.983 0.375 Colostrum & 0 h 0.001(0.000-0.000) 0.216(0.040-0.390)0.010(0.010-0.010) Bacteria 24 h 0.000(0.000-0.000) 0.012(0.000-0.020)0.085(0.010-0.160) M 0.001(0.000-0.000) 0.045(0.020-0.070) 166.60(166.60-166.60) Last 0.053(0.040-0.070) 0.000(0.000-0.000)750.00(500.0-1000.0) P value 0.143 0.276 0.100

In the model of MRSA infection, the administration of a lethal dose ofMRSA also raised IFN-7 levels to 78.13 pg/ml when treated with mixedcolostrum and bacteria. These results are similar to the IFN-7 levelsreported above for rats administered a combination of mixed colostrumand S. aureus of 78.10 pg/ml. This result compares favorably to theIFN-γ levels reported for rats administered Bovine Colostrum and S.aureus of 23.76 pg/ml. Thus, these results confirm the improved immuneresponse to treatment with mixed colostrum. The MRSA experimentalresults are summarized in Table 23 and FIG. 19. The month-long MRSAexperimental results are summarized in Table 24 and FIGS. 20A-20C.

TABLE 23 MRSA and IFN-γ Immune Response in the Presence of Colostrum(Expressed as Median pg/ml (Min-Max)) Colostrum Type: Camel Bovine Mix Pvalue^(A) Healthy  2.20(0.00-43.91) 2.20(0.00-43.91) 2.20(0.00-43.91)1.000 Colostrum 0.00(0.00-0.03)  0.09(0.00-209.50) 17.82(0.00-125.00)0.055 Bacteria 0.01(0.00-0.07) 0.57(0.01-25.05) 0.01(0.00-0.07)  0.024Colostrum & 0.00(0.00-0.87) 0.08(0.00-38.71) 78.13(0.01-300.06) 0.004Bacteria P value^(B) 0.009 0.646 0.057 ^(A)= Comparison betweendifferent treatments within a single Colostrum type using aNonparametric Test (Kruskal-Wallis Test). ^(B)= Comparison betweendifferent Colostrum types in each treatment using a Nonparametric Test(Kruskal-Wallis Test).

TABLE 24 MRSA and IFN-γ Immune Response in the Presence of Colostrum at0 h, 24 h, 7-25 days (M), and 30 days (Last) (Expressed asMedian(Min-Max)) Treatment Time Camel Bovine Mix Healthy 0 h1.496(0.220-2.770) 1.496(0.220-2.770) 1.496(0.220-2.770) 24 h3.587(1.630-5.550) 3.587(1.630-5.550) 3.587(1.630-5.550) M 42.175(40.440-43.910)  42.175(40.440-43.910)  42.175(40.440-43.910)Last 0.005(0.000-0.010) 0.005(0.000-0.010) 0.005(0.000-0.010) P value0.104 0.104 0.104 Colostrum 0 h 0.019(0.010-0.030) 1.055(0.120-1.990)3.129(0.000-6.260) 24 h 0.002(0.000-0.000) 0.058(0.060-0.060)1.667(0.000-3.330) M 0.002(0.000-0.000) 128.800(48.10-209.50)  30.327(29.390-31.260) Last 0.005(0.000-0.010) 0.001(0.000-0.000)93.750(62.50-125.00) P value 0.160 0.078 0.106 Bacteria 0 h0.043(0.020-0.070) 0.904(0.320-1.490) 0.043(0.020-0.070) 24 h0.007(0.000-0.010) 0.046(0.010-0.080) 0.007(0.000-0.010) M0.000(0.000-0.000) 12.798(0.540-25.050) 0.000(0.000-0.000) Last0.013(0.000-0.070) 1.759(0.600-2.920) 0.000(0.000-0.000) P value 0.1210.212 0.121 Colostrum & 0 h 0.600(0.330-0.870) 0.130(0.070-0.180)0.012(0.010-0.020) Bacteria 24 h 0.001(0.000-0.000) 0.056(0.020-0.090) 222.78(145.50-300.06) M 0.002(0.000-0.000) 20.951(3.190-38.710)85.565(53.92-117.21) Last 0.002(0.000-0.000) 0.009(0.000-0.010) 78.125(62.500-93.750) P value 0.104 0.104 0.112

In the model of MRSA infection, administration of camel colostrum raisedTNF-α levels to 0.56 pg/ml, compared to 0.03 pg/ml for theadministration of mixed colostrum. The greatest increase in TNF-α levelswas observed for camel colostrum, which recorded 0.69 pg/ml. Theseresults are summarized in Table 25 and FIG. 21. The month-longexperimental results are summarized in Table 26 and FIGS. 22A-22C.

TABLE 25 MRSA and TNF-α Immune Response in the Presence of Colostrum(Expressed as Median pg/ml (Min-Max)) Colostrum Type: Camel Bovine Mix Pvalue^(A) Healthy 0.00(0.00-0.60) 0.00(0.00-0.60) 0.00(0.00-0.60) 1.000Colostrum 0.56(0.00-0.94)  0.00(0.00-194.80) 0.03(0.00-0.62) 0.261Bacteria 1.17(0.50-4.15) 0.00(0.00-0.00) 1.17(0.50-4.15) 0.002 Colostrum& 0.69(0.00-4.73) 0.00(0.00-0.00)   0.08(0.00-3707.00) 0.013 Bacteria Pvalue^(b) 0.064 0.118 0.050 ^(A)= Comparison between differenttreatments within a single Colostrum type using a Nonparametric Test(Kruskal-Wallis Test). B = Comparison between different Colostrum typesin each treatment using a Nonparametric Test (Kruskal-Wallis Test).

TABLE 26 MRSA and TNF-α Immune Response in the Presence of Colostrum at0 h, 24 h, 7-25 days (M), and 30 days (Last) (Expressed asMedian(Min-Max)) Treatment Time Camel Bovine Mix Healthy 0 h0.300(0.000-0.600) 0.300(0.000-0.600) 0.300(0.000-0.600) 24 h0.220(0.000-0.440) 0.220(0.000-0.440) 0.220(0.000-0.440) M0.269(0.000-0.540) 0.269(0.000-0.540) 0.269(0.000-0.540) Last0.000(0.000-0.000) 0.000(0.000-0.000) 0.000(0.000-0.000) P value 0.6750.675 0.675 Colostrum 0 h 0.558(0.540-0.580) 0.000(0.000-0.000)0.000(0.000-0.000) 24 h 0.828(0.720-0.940) 0.000(0.000-0.000)0.000(0.000-0.000) M 0.430(0.170-0.690) 104.749(14.70-194.80) 0.177(0.050-0.300) Last 0.000(0.000-0.000) 0.000(0.000-0.000)0.490(0.360-0.620) P value 0.108 0.077 0.078 Bacteria 0 h2.884(1.620-4.150) 0.000(0.000-0.000) 2.884(1.620-4.150) 24 h0.607(0.500-0.720) 0.000(0.000-0.000) 0.607(0.500-0.720) M0.000(0.000-0.000) 0.000(0.000-0.000) 0.000(0.000-0.000) Last1.166(0.500-4.150) 0.000(0.000-0.000) 0.000(0.000-0.000) P value 0.1211.000 0.121 Colostrum & 0 h 4.098(3.470-4.730) 0.000(0.000-0.000)0.000(0.000-0.000) Bacteria 24 h 1.422(0.940-1.910) 0.000(0.000-0.000)0.000(0.000-0.000) M 0.383(0.330-0.440) 0.000(0.000-0.000)1853.58(0.16-3707.00)  Last 0.000(0.000-0.000) 0.000(0.000-0.000)0.622(0.560-0.680) P value 0.080 1.000 0.109

In the model of MRSA infection, administration of mixed colostrum raisedIL-10 levels during the extended testing period to similar levelsobserved for S. aureus. Moreover, the administration of bovine colostrumincreased IL-10 to 0.810 pg/ml after 24 hours. These results aresummarized in Table 27 and FIG. 23. The month-long experimental resultsare summarized in Table 28 and FIGS. 24A-24C.

TABLE 27 MRSA and IL-10 Immune Response in the Presence of Colostrum(Expressed as Median pg/ml (Min-Max)) Colostrum Type: Camel Bovine Mix Pvalue^(A) Healthy 0.00(0.00-36.76) 0.00(0.00-36.76) 0.00(0.00-36.76) 1.000 Colostrum 0.00(0.00-16.30) 0.14(0.00-38.65) 83.30(0.00-1000.00)0.115 Bacteria 0.00(0.00-0.00)  0.04(0.02-0.34)  0.00(0.00-0.00)  0.001Colostrum & 0.00(0.00-0.04)  0.03(0.00-6.41)  0.01(0.00-500.00) 0.035Bacteria P value^(B) 0.057 0.348 0.002 ^(A)= Comparison betweendifferent treatments within a single Colostrum type using aNonparametric Test (Kruskal-Wallis Test). ^(B)= Comparison betweendifferent Colostrum types in each treatment using a Nonparametric Test(Kruskal-Wallis Test).

TABLE 28 MRSA and IL-10 Immune Response in the Presence of Colostrum at0 h, 24 h, 7-25 days (M), and 30 days (Last) (Expressed asMedian(Min-Max)) Treatment Time Camel Bovine Mix Healthy 0 h0.016(0.000-0.030) 0.016(0.000-0.030) 0.016(0.000-0.030) 24 h18.378(0.000-36.760) 18.378(0.000-36.760) 18.378(0.000-36.760) M0.018(0.000-0.030) 0.018(0.000-0.030) 0.018(0.000-0.030) Last0.004(0.000-0.010) 0.004(0.000-0.010) 0.004(0.000-0.010) P value 0.9350.935 0.935 Colostrum 0 h 0.000(0.000-0.000) 19.621(0.590-38.650)0.001(0.000-0.000) 24 h 0.000(0.000-0.000) 0.810(0.150-1.470)0.002(0.000-0.000) M  8.152(0.000-16.300) 0.077(0.020-0.130) 166.63(166.60-166.66) Last 0.005(0.000-0.000) 0.001(0.000-0.000)750.00(500.0-1000.0) P value 0.105 0.104 0.091 Bacteria 0 h0.000(0.000-0.000) 0.207(0.070-0.340) 0.000(0.000-0.000) 24 h0.000(0.000-0.000) 0.024(0.020-0.020) 0.000(0.000-0.000) M0.000(0.000-0.000) 0.037(0.030-0.050) 0.000(0.000-0.000) Last0.000(0.000-0.000) 0.036(0.030-0.040) 0.000(0.000-0.000) P value 1.0000.108 1.000 Colostrum & 0 h 0.019(0.000-0.040) 1.089(0.030-2.150)0.008(0.000-0.010) Bacteria 24 h 0.000(0.000-0.000) 3.225(0.040-6.410)0.006(0.000-0.010) M 0.001(0.000-0.000) 0.034(0.020-0.040)0.006(0.000-0.010) Last 0.003(0.000-0.000) 0.002(0.000-0.000) 500.00(500.00-500.00) P value 0.392 0.198 0.222

Overall, the S. aureus lethal dose experiments confirm that treatmentwith bovine colostrum resulted in lower stimulation of immune systemcytokines than other tested treatments. For example, both theadministration of colostrum mix alone and the administration ofcolostrum mix and the lethal dose of bacteria resulted in the sameincrease in IL-10 levels (83.30 pg/ml compared to administration ofbacteria alone at 0.07 pg/ml) Administration of mixed colostrum aloneand administration of mixed colostrum and bacteria raised IL-10 levelsto 166.63 pg/ml at days 7-25 (M) and to 750.00 pg/ml by the final day ofthe experiment. Both camel colostrum and mixed camel colostrum andbovine colostrum stimulated TNF-α levels. The most enormous change wasfor camel colostrum in all treatments, which resulted in 0.56 pg/ml,0.50 pg/ml, and 1.21 pg/ml for colostrum alone, bacteria alone, andcolostrum and bacteria respectively.

Example 5 Calculating Correlations Between the Studied Cytokines and theColostrum Sources

The immunomodulatory effect of pro-inflammatory cytokines varieddepending upon the bacterial species tested. IFN-γ exhibited a steadylevel of immune response in mixed colostrum treatment (93.750 pg/ml inthe E. coli model). In the S. aureus model, IFN-γ levels increased to78.10 pg/ml when the mixture of colostrum was administered compared to17.82 pg/ml in the control colostrum. In the MRSA experiments, IFN-γlevels increased to 78.13 pg/ml when the mixed colostrum wasadministered. TNF-α, in the P. aeruginosa experiments showed the mostconsiderable change in level when camel colostrum and bacteria wereadministered (8.931 pg/ml after 24 hours compared to 1.171 pg/ml forbacterial injection only).

IL-10 is a crucial cytokine marker. In E. coli treated with mixedcolostrum, IL-10 was elevated to 83.30 pg/ml compared to 31.62 pg/mlwhen bacteria alone were administered. Early cytokine transcriptionalchanges could be useful as a forecasting tool. IL-10 started to increaseafter 24 hours in the E. coli experiments, up to 80.3080 pg/ml. In S.aureus administered mixed colostrum and bacteria, IL-10 levels rose to166.60 pg/ml after 7-25 days, and this increase was sustained over timeand ended with 750.00 pg/ml after one month.

The best survival effect was in camel colostrum and mixed colostrum forall the tested bacteria (83% in E-coli-100% in MRSA) in the infectedrats in the presence of colostrum.

Camel colostrum and mixed colostrum blocked adherence of bacteria tocultured tissue cells. Mixed colostrum (bovine colostrum and camelcolostrum) can stimulate the immune response more than the bovinecolostrum or camel colostrum alone. Colostrum has been shown to be aconstant source of potentially probiotic bacteria in an infant's gut,including staphylococci, streptococci, and lactic acid bacteria. Theimmunomodulatory effect of pro-inflammatory cytokines varied due to thebacterial species.

IL-10 and IFN-γ showed a significant interaction at 24 hours. Thesignificant elevations of the IL-10 and TNF-α transcriptional levelsmost likely indicate their essential role in the regulation of theimmune responses of the bovine mammary gland in S. aureus infection.That could reflect the suppressive nature of the S. aureus mastitis.Early cytokine transcriptional changes could be useful as a forecastingtool in the detection of the subclinical S. aureus mastitis. Thetranscriptional patterns of these cytokines in the early stages of S.aureus infection could help to unravel their role in the pathophysiologyof S aureus mastitis. Finally, vital, crucial cytokine marker(s) couldemerge that provide efficient diagnostic and therapeutic means for theearly detection of S. aureus mastitis. Early cytokine transcriptionalchanges could be useful as a forecasting tool in the detection of thesubclinical S. aureus mastitis.

These experiments demonstrate that camel colostrum, bovine colostrum andmixed colostrum (bovine and camel colostrum) can provide protective andimmune-stimulatory effects against the studied pathogenic bacteria invitro and in vivo. The lyophilized colostrum derivative studiedmaintained most of the bioactive factors that appear to be at the basisof its many nutritional, modulatory, or even therapeutic, applications.

The fastest and the best-protected treatment was for lethal S. aureuswith a baseline of 50-100% mortality. 100% survival was obtained for allsix rats injected with S. aureus and administered the three types ofcolostrum by i.p. injection. Camel colostrum had a tremendous protectiveeffect from lethal MRSA, demonstrating 100% survival versus 100%mortality in MRSA administration alone. Also, MRSA showed the highestmaximum increased values in % change in weight of rats administeredmixed colostrum (43% compared to −21% for rats administered only thelethal dose of MRSA).

Camel colostrum and mixed colostrum alone induced a remarkable increasein the weight of rats ranging from (6%-59%). Bacterial injection led toa dramatic decrease in the percent weight change in rats from −12% to12%. MRSA demonstrated the most significant maximum increased values inpercent change of weight of rats treated with mixed colostrum (43%compared to −21% when administered MRSA alone).

Calculations of the correlation between IFN-γ, IL-10, and TNF-α aresummarized in Tables 29-31. Table 29 shows that without the presence ofColostrum, the value of the correlation coefficient between IL-10 andTNF-α was −0.773**, indicating a significant negative strength ofassociation, while the value of the correlation coefficient betweenIFN-γ and TNF-α was 0.464*, demonstrating a less significant positivestrength of association. With the presence of Colostrum, the onlysignificant value of the correlation coefficient was 0.647**, betweenIL-10 and IFN-γ. In the absence of Colostrum, the administration ofbacterial strains did not demonstrate a significant value of thecorrelation coefficient between the three tested cytokines in gramnegative E. coli and P. aeruginosa; however, gram-positive S. aureus andMRSA has significant values of the correlation coefficient between IL-10and IFN-γ. In the presence of both Colostrum and bacterial strains, thecorrelation coefficient between IL-10 and IFN-γ demonstrated a positivestrength of association for S. aureus (0.701*) and MRSA (0.503*).Further, in the presence of E. coli, the correlation coefficient betweenIL-10 and TNF-α had a significant negative strength of association(−0.470*).

Table 30 shows the correlation between IFN-γ, IL-10, and TNF-α in thepresence of Colostrum when treated with S. aureus or MRSA. A significantvalue of a correlation coefficient was observed in the presence of CamelColostrum, with the coefficient between IL-10 and TNF-α having anegative strength of association for S. aureus of −0.517** and for MRSAof −0.585**. A significant value of a correlation coefficient wasobserved in the presence of Bovine Colostrum, with the coefficientbetween IFN-γ and TNF-α having a positive strength of association for S.aureus of 0.415* and for MRSA of 0.551**. A number of significant valuesof correlation coefficients were observed in the presence of Mixed Cameland Bovine Colostrum, with the coefficient between IFN-γ and IL-10,IFN-γ and TNF-α, and IL-10 and TNF-α all having respective positivestrength of association for S. aureus of 0.659**, 0.694**, and 0.439*.For MRSA, the coefficient between IFN-γ and IL-10 also showed a positivestrength of association of 0.512**.

TABLE 29 Correlation between IFN-γ, IL-10, and TNF-α R (SpearmanTreatments Parameters Correlation) Sig. Healthy IFN-γ with IL10 −0.0360.867 N^(b) IFN-γ with TNF-α 0.464* 0.022 P^(a) IL10 with TNF-α −0.773**0.000 N^(b) Colostrum IFN-γ with IL10 0.647** 0.001 P^(a) IFN-γ withTNF-α 0.269 0.203 P^(a) IL10 with TNF-α 0.025 0.908 P^(a) E. coli IFN-γwith IL10 0.242 0.267 P^(a) IFN-γ with TNF-α 0.056 0.811 P^(a) IL10 withTNF-α −0.295 0.207 N^(b) Colostrum & IFN-γ with IL10 0.353 0.151 P^(a)E. coli IFN-γ with TNF-α −0.144 0.557 N^(b) IL10 with TNF-α −0.470*0.049 N^(b) P. aeruginosa IFN-γ with IL10 0.490 0.054 P^(a) IFN-γ withTNF-α −0.226 0.384 N^(b) IL10 with TNF-α −0.329 0.214 N^(b) Colostrum &IFN-γ with IL10 0.186 0.418 P^(a) P. aeruginosa IFN-γ with TNF-α −0.0820.732 N^(b) IL10 with TNF-α 0.150 0.528 P^(a) S. aureus IFN-γ with IL100.755** 0.000 P^(a) IFN-γ with TNF-α −0.168 0.443 N^(b) IL10 with TNF-α−0.278 0.211 N^(b) Colostrum & IFN-γ with IL10 0.701** 0.000 P^(a) S.aureus IFN-γ with TNF-α 0.061 0.782 P^(a) IL10 with TNF-α −0.135 0.550N^(b) MRSA IFN-γ with IL10 0.796** 0.000 P^(a) IFN-γ with TNF-α −0.512*0.043 N^(b) IL10 with TNF-α −0.863** 0.000 N^(b) Colostrum & IFN-γ withIL10 0.503* 0.014 P^(a) MRSA IFN-γ with TNF-α 0.009 0.968 P^(a) IL10with TNF-α −0.288 0.182 N^(b) *= Correlation is significant at the 0.05level. **= Correlation is significant at the 0.01 level. ^(a)= Positivecorrelation. ^(b)= Negative correlation.

TABLE 30 Correlation between IFN-γ, IL-10, and TNF-α in the presence ofcolostrum for S. aureus and MRSA R (Spearman Colostrum Type ParametersCorrelation) Sig. S. aureus Camel IFN-γ with IL10 0.140 0.452 P^(a)IFN-γ with TNF-α 0.090 0.628 P^(a) IL10 with TNF-α −0.517** 0.003 N^(b)Bovine IFN-γ with IL10 0.242 0.207 P^(a) IFN-γ with TNF-α 0.415* 0.020P^(a) IL10 with TNF-α −0.290 0.121 N^(b) Mix IFN-γ with IL10 0.659**0.000 P^(a) IFN-γ with TNF-α 0.694** 0.000 P^(a) IL10 with TNF-α 0.439*0.013 P^(a) MRSA Camel IFN-y with IL10 0.116 0.566 P^(a) IFN-y withTNF-α 0.030 0.880 P^(a) IL10 with TNF-α −0.585** 0.001 N^(b) BovineIFN-y with IL10 0.216 0.236 P^(a) IFN-y with TNF-α 0.551** 0.001 P^(a)IL10 with TNF-α −0.325 0.070 N^(b) Mix IFN-y with IL10 0.512** 0.005P^(a) IFN-y with TNF-α 0.221 0.258 P^(a) IL10 with TNF-α −0.219 0.264N^(b) *= Correlation is significant at the 0.05 level. **= Correlationis significant at the 0.01 level. ^(a)= Positive correlation. ^(b)=Negative correlation.

Table 31 shows the correlation between IFN-γ, IL-10, and TNF-α in thepresence of Colostrum when treated with E. coli or P. aeruginosa. Asignificant value of a correlation coefficient was observed in thepresence of Camel Colostrum, with the coefficient between IL-10 andTNF-α having a negative strength of association for E. coli of −0.595**.A significant value of a correlation coefficient was observed in thepresence of Bovine Colostrum, with the coefficient between IFN-γ andTNF-α having a positive strength of association for E. coli of 0.559**and for P. aeruginosa of 0.548**. A number of significant values ofcorrelation coefficients were observed in the presence of Mixed Cameland Bovine Colostrum, with the coefficient between IFN-γ and TNF-αhaving a positive strength of association for E. coli of 0.625** and forP. aeruginosa of 0.739**. Further, significant values of correlationcoefficients were observed in the presence of Mixed Camel and BovineColostrum, with the coefficient between IFN-7 and IL-10 also showing apositive strength of association of 0.412* for E. coli and of 0.374* forP. aeruginosa.

TABLE 31 Correlation between IFN-γ, IL-10, and TNF-α in the presence ofcolostrum for E. coli and P. aeruginosa R (Spearman Colostrum TypeParameters Correlation) Sig. E. coli Camel IFN-γ with IL10 0.078 0.688P^(a) IFN-γ with TNF-α −0.203 0.282 N^(b) IL10 with TNF-α −0.595** 0.001N^(b) Bovine IFN-γ with IL10 0.345 0.057 P^(a) IFN-γ with TNF-α 0.559**0.002 P^(a) IL10 with TNF-α −0.209 0.287 N^(b) Mix IFN-γ with IL100.413* 0.026 P^(a) IFN-γ with TNF-α 0.625** 0.000 P^(a) IL10 with TNF-α0.210 0.274 P^(a) P. aeruginosa Camel IFN-γ with IL10 0.088 0.664 P^(a)IFN-γ with TNF-α −0.120 0.550 N^(b) IL10 with TNF-α −0.043 0.832 N^(b)Bovine IFN-γ with IL10 0.198 0.323 P^(a) IFN-γ with TNF-α 0.548** 0.003P^(a) IL10 with TNF-α −0.277 0.162 N^(b) Mix IFN-γ with IL10 0.374*0.038 P^(a) IFN-γ with TNF-α 0.739** 0.000 P^(a) IL10 with TNF-α 0.2140.257 P^(a) *= Correlation is significant at the 0.05 level. **=Correlation is significant at the 0.01 level. ^(a)= Positivecorrelation. ^(b)= Negative correlation.

Overall, these examples illustrate at least that mixed colostrum cantrigger a greater immune response than the administration of eitherbovine colostrum or camel colostrum alone. Further, the administrationof any of the three colostrum groups increased survival against lethaldoses of gram-positive and gram-negative bacteria, with camel colostrumand mixed colostrum providing the greatest increase in survival.

It is to be understood that the methods described herein are not limitedto the specific embodiments described above, but encompasses any and allembodiments within the scope of the generic language of the followingclaims enabled by the embodiments described herein, or otherwise shownin the drawings or described above in terms sufficient to enable one ofordinary skill in the art to make and use the claimed subject matter.

1-18. (canceled)
 19. A colostrum composition comprising: a lyophilizedcolostrum, the lyophilized colostrum being made by the steps of:collecting colostrum from the mammary gland of a mammal; freezing thecolostrum at a temperature of at least −20° C. for at least a week;thawing the colostrum for at least 8 hours; lyophilizing the colostrum;and suspending the lyophilized colostrum in a saline solution to providea colostrum composition.
 20. A natural pharmaceutical compositioncomprising the colostrum composition of claim 19 and a pharmaceuticallyacceptable carrier.